CN117462551A

CN117462551A - Application of indinavir and/or derivative thereof in preparation of antituberculosis drugs

Info

Publication number: CN117462551A
Application number: CN202311811103.1A
Authority: CN
Inventors: 郭庆龙; 毕静; 张国良
Original assignee: Shenzhen National Clinical Research Center For Infectious Diseases
Current assignee: Shenzhen National Clinical Research Center For Infectious Diseases
Priority date: 2023-12-27
Filing date: 2023-12-27
Publication date: 2024-01-30

Abstract

The application provides application of indinavir and/or derivatives thereof in preparing antituberculosis drugs, and relates to the technical field of medicines. Indinavir is an antiretroproteinase inhibitor, can be combined with HIV protease catalytic sites, and can inhibit the division of virus polyprotein precursor into mature functional proteins required for virus replication. Indinavir can be used with other anti-reverse transcription virus medicaments to treat HIV infection; the application finds that indinavir has the effect of inhibiting the intracellular activity of tubercle bacillus at the level of macrophages, namely the indinavir has the potential of being used as an antitubercular drug, and has important application value in the clinical treatment of HIV combined MTB infection.

Description

Application of indinavir and/or derivative thereof in preparation of antituberculosis drugs

Technical Field

The application relates to the technical field of medicines, in particular to application of indinavir and/or derivatives thereof in preparation of antituberculosis drugs.

Background

AIDS (acquired immunodeficiency syndrome, AIDS) is an important public health problem threatening public health.

AIDS patients are immunodeficient and often combine multiple opportunistic infections, while mycobacterium tuberculosis (Mycobacterium tuberculosis, MTB) is the more common pathogen and progresses more frequently, with only HIV infection combining latent mycobacterium tuberculosis infection (Latent TB Infection, LTBI) progressing to active tuberculosis 30 times more than HIV negative. Thus, HIV interacts with, and affects, MTB, and diagnosis is complex and difficult. Early identification and discovery of tuberculosis (including LTBI and active tuberculosis), and formulation of effective anti-tuberculosis and antiviral therapy (anti-retroviral therapy, ART) regimens are the most effective key measures to reduce the risk of HIV/MTB double infection; in the related art, no medicine with the functions of preventing, controlling and promoting curative effect on HIV/MTB double infection exists.

Disclosure of Invention

The present application has been made in view of the above problems, and an object thereof is to provide an application of indinavir and/or its derivatives in preparing antitubercular drugs; the medicine can prevent and control HIV/MTB double infection.

The first aspect of the present application provides the use of indinavir and/or its derivatives in the preparation of antitubercular medicaments.

Indinavir is an antiretroproteinase inhibitor, can be combined with HIV protease catalytic sites, and can inhibit the division of virus polyprotein precursor into mature functional proteins required for virus replication. The molecular formula is as follows:

indinavir can be used with other anti-reverse transcription virus medicaments to treat HIV infection; the application finds that indinavir has the effect of inhibiting the intracellular activity of tubercle bacillus at the level of macrophages, namely the indinavir has the potential of being used as an antitubercular drug, and has important application value in the clinical treatment of HIV combined MTB infection.

Optionally, the derivative of indinavir includes pharmaceutically acceptable salts thereof.

Alternatively, the pharmacologically acceptable salts include salts with inorganic acids, organic acids, alkali metals, alkaline earth metals, and amino acids.

Optionally, the antitubercular drug is used for preventing or treating infection by tubercle bacillus.

Optionally, the preparation raw materials of the antituberculosis drug also comprise a medicinal carrier.

Optionally, the pharmaceutically acceptable carrier comprises at least one of diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, sweeteners, and flavoring agents.

In a second aspect, the present application provides a medicament for preventing and/or treating tuberculosis, the active ingredient of the medicament comprises indinavir and/or derivatives thereof.

Optionally, the mass content of indinavir and/or derivative thereof in the medicine is 0.1% -99%.

Optionally, the tuberculosis comprises pulmonary tuberculosis or extrapulmonary tuberculosis.

Optionally, the drug is administered in at least one form selected from the group consisting of powders, tablets, granules, capsules, solutions, emulsions, and suspensions.

In a third aspect, the present application provides a tuberculosis treatment kit comprising the above-described medicament for preventing and/or treating tuberculosis.

Optionally, the content of indinavir and/or derivative thereof in the kit is formulated according to the following daily dosage standard:

indinavir and/or derivatives thereof: 0.1mg/kg body weight/day to 1000mg/kg body weight/day.

Drawings

In order to more clearly illustrate the embodiments of the present drawings or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present drawings, and that other drawings may be obtained according to the structures shown in these drawings without inventive effort to a person skilled in the art.

FIG. 1 is a graph showing the effect of various concentrations of indinavir on the cell viability of THP-1 macrophages in example 1.

FIG. 2 is a graph showing the results of inhibition of Mycobacterium tuberculosis H37Rv by indinavir at macrophage level in example 2.

The achievement of the objects, functional features and advantages of the present drawings will be further described with reference to the accompanying drawings in conjunction with the embodiments.

Detailed Description

Hereinafter, embodiments of the application of indinavir and/or its derivatives in the preparation of antitubercular drugs of the present application are specifically disclosed with reference to the accompanying drawings as appropriate. However, unnecessary detailed description may be omitted. For example, detailed descriptions of well-known matters and repeated descriptions of the actual same structure may be omitted. This is to avoid that the following description becomes unnecessarily lengthy, facilitating the understanding of those skilled in the art. Furthermore, the drawings and the following description are provided for a full understanding of the present application by those skilled in the art, and are not intended to limit the subject matter recited in the claims.

The "range" disclosed herein is defined in terms of lower and upper limits, with a given range being defined by the selection of a lower and an upper limit, the selected lower and upper limits defining the boundaries of the particular range. Ranges that are defined in this way can be inclusive or exclusive of the endpoints, and any combination can be made, i.e., any lower limit can be combined with any upper limit to form a range. For example, if ranges of 60-120 and 80-110 are listed for a particular parameter, it is understood that ranges of 60-110 and 80-120 are also contemplated. Furthermore, if the minimum range values 1 and 2 are listed, and if the maximum range values 3,4 and 5 are listed, the following ranges are all contemplated: 1-3, 1-4, 1-5, 2-3, 2-4 and 2-5. In this application, unless otherwise indicated, the numerical range "a-b" represents a shorthand representation of any combination of real numbers between a and b, where a and b are both real numbers. For example, the numerical range "0-5" means that all real numbers between "0-5" have been listed throughout, and "0-5" is simply a shorthand representation of a combination of these values. When a certain parameter is expressed as an integer of 2 or more, it is disclosed that the parameter is, for example, an integer of 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12 or the like.

All embodiments and alternative embodiments of the present application may be combined with each other to form new solutions, unless specifically stated otherwise.

All technical features and optional technical features of the present application may be combined with each other to form new technical solutions, unless specified otherwise.

All steps of the present application may be performed sequentially or randomly, preferably sequentially, unless otherwise indicated. For example, the method comprises steps (a) and (b), meaning that the method may comprise steps (a) and (b) performed sequentially, or may comprise steps (b) and (a) performed sequentially. For example, the method may further include step (c), which means that step (c) may be added to the method in any order, for example, the method may include steps (a), (b) and (c), may include steps (a), (c) and (b), may include steps (c), (a) and (b), and the like.

Reference herein to "comprising" and "including" means open ended, as well as closed ended, unless otherwise noted. For example, the terms "comprising" and "comprises" may mean that other components not listed may be included or included, or that only listed components may be included or included.

The term "or" is inclusive in this application, unless otherwise specified. For example, the phrase "a or B" means "a, B, or both a and B. More specifically, either of the following conditions satisfies the condition "a or B": a is true (or present) and B is false (or absent); a is false (or absent) and B is true (or present); or both A and B are true (or present).

Tuberculosis is a chronic infectious disease caused by a mycobacterium tuberculosis complex (Mycobacterium tuberculosis complex, abbreviated as mycobacterium tuberculosis or tubercle bacillus) and can involve a whole body multi-organ system, and the most common diseased part is the lung, which accounts for 80% -90% of the total number of tuberculosis of each organ. Mycobacterium tuberculosis complex includes Mycobacterium tuberculosis (Mycobacterium tuberculosis, MTB), mycobacterium bovis, mycobacterium africanum and Mycobacterium vaccae, the main causing human diseases are Mycobacterium tuberculosis.

AIDS patients are immunodeficient and often combine multiple opportunistic infections, while mycobacterium tuberculosis is the more common pathogen and progresses more frequently, with only HIV infection combining with a latent mycobacterium tuberculosis infected person who progresses to active tuberculosis 30 times more than HIV negative. Thus, HIV interacts with, and affects, MTB, and diagnosis is complex and difficult. Therefore, controlling the spread of tuberculosis, preventing new infection by HIV, and standardizing the diagnosis and treatment of double-infected persons are important works and challenges in disease prevention and control. In the related art, no medicine with the functions of preventing, controlling and promoting curative effect on HIV/MTB double infection exists.

For this purpose, a first aspect of the embodiments of the present application provides the use of indinavir and/or its derivatives in the preparation of antitubercular drugs.

Indinavir is a non-nucleoside reverse transcriptase inhibitor, and has the molecular formula:

；

indinavir is directly combined with HIV-1 reverse transcriptase, and blocks RNA-dependent and DNA-dependent DNA polymerase activity by destroying enzyme catalytic sites, so that indinavir is a medicine for treating HIV. The embodiment of the application finds that indinavir has the effect of inhibiting the intracellular survival of tubercle bacillus at the level of macrophages, and has the potential of being used as an antitubercular drug, and has important application value in the clinical treatment of HIV combined MTB infection.

In some embodiments, the derivative of indinavir includes pharmaceutically acceptable salts thereof.

In some embodiments, the pharmacologically acceptable salts include salts with inorganic acids, organic acids, alkali metals, alkaline earth metals, and amino acids.

In some embodiments, the pharmacologically acceptable salts described herein include salts with inorganic acids, organic acids, alkali metals, alkaline earth metals, and amino acids.

In some embodiments, the inorganic acid comprises at least one of hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, hydrobromic acid.

In some embodiments, the organic acid comprises at least one of maleic acid, fumaric acid, tartaric acid, lactic acid, citric acid, acetic acid, methanesulfonic acid, p-toluenesulfonic acid, adipic acid, palmitic acid, and tannic acid.

In some embodiments, the alkali metal comprises at least one of lithium, sodium, and potassium.

In some embodiments, the alkaline earth metal comprises at least one of calcium and magnesium.

In some embodiments, the amino acid comprises lysine.

In some embodiments, the antitubercular drug is used to prevent or treat infection by mycobacterium tuberculosis.

Mycobacterium tuberculosis, also known as Mycobacterium tuberculosis, is a causative agent of tuberculosis. Mycobacterium tuberculosis H37Rv is a strain isolated in 1905 and widely used in biomedical research worldwide, and has an animal model of tuberculosis with complete toxicity.

In some embodiments, the preventing or treating an infection by a tubercle bacillus specifically refers to killing the tubercle bacillus or inhibiting the growth and reproduction of the tubercle bacillus.

In some embodiments, the antitubercular drug formulation further comprises a pharmaceutically acceptable carrier.

In some embodiments, the pharmaceutically acceptable carrier comprises at least one of diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, sweeteners, and flavoring agents.

In some embodiments, the excipient comprises at least one of lactose, polyethylene glycol, sodium citrate, dicalcium phosphate, and water.

In some embodiments, the filler comprises at least one of starch, lactose, sucrose, glucose, mannitol, and silicic acid.

In some embodiments, the binder comprises at least one of cellulose derivatives, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic.

In some embodiments, the cellulose derivative comprises carboxymethyl cellulose.

In some embodiments, the alginate comprises alginate.

In some embodiments, the humectant comprises glycerin.

In some embodiments, the disintegrant comprises at least one of agar-agar, calcium carbonate, sodium bicarbonate, potato starch, tapioca starch, alginic acid, silicate, and sodium carbonate.

In some embodiments, the absorption enhancer comprises a quaternary ammonium compound.

In some embodiments, the surfactant comprises cetyl alcohol.

In some embodiments, the adsorption carrier comprises at least one of kaolin clay, bentonite clay, and a soap clay.

In some embodiments, the lubricant comprises at least one of talc, calcium stearate, magnesium stearate, sodium lauryl sulfate, and polyethylene glycol.

In some embodiments, the dosage form of the medicament is a variety of dosage forms conventional in the art.

In some embodiments, the mass content of indinavir and/or its derivatives in the medicament for preventing and/or treating tuberculosis is 0.1% -99%.

In some embodiments, the mass content of indinavir and/or its derivatives in the medicament for preventing and/or treating tuberculosis is 0.1% -95%.

In some embodiments, the tuberculosis refers to various types of tuberculosis that mycobacterium tuberculosis can cause.

In some embodiments, the tuberculosis comprises tuberculosis or extrapulmonary tuberculosis.

In some embodiments, the extrapulmonary tuberculosis comprises at least one of a bone joint tuberculosis, tuberculous meningitis, tuberculous pleurisy, renal tuberculosis, and intestinal tuberculosis.

In some embodiments, the medicament for preventing and/or treating tuberculosis may be prepared in a variety of dosage forms for ease of administration.

In some embodiments, the pharmaceutical dosage form for preventing and/or treating tuberculosis is at least one of powder, tablet, capsule, soft capsule, granule, pill, oral liquid, dry suspension, drop pill, dry extract, injection and infusion.

In some embodiments, the mode of administration of the drug for preventing and/or treating tuberculosis may be a mode of administration conventional in the art, including, but not limited to, injection or oral administration.

In some embodiments, the administration by injection may be intravenous, intramuscular, intraperitoneal, intradermal, or subcutaneous injection.

In some embodiments, the agent for preventing and/or treating tuberculosis is administered orally, rectally, parenterally, intravaginally, intraperitoneally, topically, bucally, to humans and other mammals, or as an oral spray or nasal spray.

In some embodiments, the topical comprises administration to humans and other mammals by powder, ointment or drops.

The term "parenteral" as used herein refers to modes of administration including intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intra-articular injection and infusion.

In some embodiments, the medicament for preventing and/or treating tuberculosis may be an oral formulation in particular, so as to be orally administered.

In some embodiments, the oral formulation is specifically a solid dosage form or a liquid dosage form; optionally, the solid dosage form is in particular a tablet, dragee, pill, powder, granule, capsule or coating; alternatively, the liquid dosage form may be, in particular, an emulsion, solution, suspension, syrup or elixir.

In some embodiments, the capsule, tablet or pill further comprises a buffer.

In some embodiments, the solid dosage forms of tablets, dragees, capsules, pills, and granules may be prepared with coatings and shells; optionally, the shell material includes an enteric coating material and other coating materials known in the pharmaceutical formulation arts.

In some embodiments, the solid dosage form incorporates an opacifying agent.

In some embodiments, the composition of the agent for preventing and/or treating tuberculosis may also be such that it releases the active ingredient only at a certain part of the intestinal tract or preferentially at a certain part of the intestinal tract in a delayed manner.

In some embodiments, the drug for preventing and/or treating tuberculosis is prepared in the form of an embedding composition using a polymer or wax.

In some embodiments, the drug for preventing and/or treating tuberculosis is formulated in a microcapsule form.

In some embodiments, the liquid dosage form in the oral formulation includes a diluent or solubilizer in addition to the drug for preventing and/or treating tuberculosis.

In some embodiments, the diluent comprises water or an organic solvent.

In some embodiments, the solubilizing agent comprises at least one of ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, dimethylformamide, vegetable oils, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols, and fatty acid esters of sorbitan.

In some embodiments, the vegetable oil comprises at least one of cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil.

In some embodiments, the medicament may be specifically an parenteral formulation.

In some embodiments, the parenteral formulation comprises an injection or a suppository.

In some embodiments, the injection is a solution, suspension, or injectable dry powder.

Injectable dry powders refer to the addition of injection water immediately prior to injection.

In some embodiments, the injection comprises, in addition to a medicament for preventing and/or treating tuberculosis, the following carriers or adjuvants: a physiologically acceptable sterile aqueous solution, non-aqueous solution, dispersion, suspension, emulsion, aqueous carrier, non-aqueous carrier, diluent, solvent or vehicle, such as, in particular, at least one of water, ethanol, polyol, vegetable oil and injectable organic ester.

In some embodiments, the polyol includes at least one of propylene glycol, polyethylene glycol, and glycerin.

In some embodiments, the vegetable oil comprises olive oil.

In some embodiments, the injectable organic ester comprises ethyl oleate.

In some embodiments, the injection may also be in the form of an injectable depot formulation.

In some embodiments, the injectable depot formulation can be prepared by forming a microcapsule matrix of the drug in a biodegradable polymer.

In some embodiments, the biodegradable polymer comprises polylactide-polyglycolide, polyorthoester, or polyanhydrides.

In some embodiments, the injectable depot formulation is capable of controlling the rate of release of the drug by the ratio of the drug to the polymer and the nature of the particular polymer employed.

In some embodiments, the injectable depot formulations may also be prepared by embedding the drug in liposomes or microemulsions which are compatible with body tissues.

In some embodiments, the injectable formulation is further subjected to a sterilization process.

In some embodiments, the sterilization process includes filtering or incorporating the sterilant in the form of a sterile solid composition with a bacteria filter.

In some embodiments, the sterile solid composition may be dissolved or dispersed in sterile water or other sterile injectable medium just prior to use.

In some embodiments, suspending agents may also be included in the injectable and oral suspensions.

In some embodiments, the suspending agent comprises at least one of ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, polyoxyethylene sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth.

Furthermore, in some cases, it is desirable to slow down the absorption of subcutaneously or intramuscularly injected drugs in order to prolong the effect of the drugs. This can be achieved by using liquid suspensions of crystalline or amorphous materials that are poorly water soluble; the absorption rate of the drug depends on the dissolution rate, which in turn depends on the crystal size and the crystal form, and the purpose of prolonging the drug action can be achieved by adding the above substances. In addition, delayed absorption of parenterally administered pharmaceutical forms may be accomplished by dissolving or suspending the drug in an oil vehicle.

In some embodiments, the suppositories may be used for rectal or vaginal administration; the suppositories can be prepared by mixing the drugs for preventing and/or treating tuberculosis of the present example with a suitable non-irritating excipient or carrier; the excipient or carrier is particularly preferably cocoa butter, polyethylene glycol or a suppository wax; the excipient or carrier is solid at room temperature, but liquid at the body temperature of a human or other mammal, and thus melts in the rectal or vaginal cavity to release the active compound.

In some embodiments, the medicament for preventing and/or treating tuberculosis may be a topical administration formulation, such as a powder, a spray, an ointment, and an inhalant.

Mixing the active ingredient (indinavir and/or derivatives thereof) in the medicine for preventing and/or treating tuberculosis with pharmaceutically acceptable carrier, required preservative, buffer and propellant under aseptic condition to obtain topical administration preparation; in addition, ophthalmic formulations, eye ointments, powders and solutions are also contemplated as being within the scope of the examples.

In some embodiments, the indinavir and/or derivative thereof can be specifically a liposomal formulation, administered in liposomal form. The specific preparation method of the liposome preparation is not particularly limited in the examples, and methods well known to those skilled in the art may be used.

Liposomes are commonly made in the art with phospholipids or other lipid substances; liposomes are formed from a single or multiple layers of hydrated liquid crystals dispersed in an aqueous medium, and any non-toxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used. In some embodiments, the lipid is a natural and synthetic phospholipid and/or phosphatidylcholine (lecithin). In addition, the liposome-form preparation may contain a stabilizer, a preservative, an excipient, etc., in addition to the active ingredient in the drug for preventing and/or treating tuberculosis of the present application.

The preparation method of the dosage form according to the scheme is not particularly required, and the method known by the person skilled in the art is used for preparing the active ingredient in the drug for preventing and/or treating tuberculosis provided by the application into the drug with the corresponding dosage form.

The medicament for preventing and/or treating tuberculosis of the present application can be used for preventing or treating tuberculosis of mammals (such as human beings), and a subject in need thereof is provided with a prophylactically or therapeutically effective amount of the medicament for preventing and/or treating tuberculosis. The medicament for preventing and/or treating tuberculosis provided by the application can be applied in a pure form, or applied in a prodrug form, or applied in a medicament form containing the medicament for preventing and/or treating tuberculosis provided by the application.

In particular embodiments of the present application, the actual dosage level of the active ingredient (indinavir and/or derivatives thereof) in the medicaments of the present application can be varied to provide an amount of active compound effective to achieve the desired therapeutic response for a particular patient, composition and mode of administration. The dosage level will be selected based on the activity of the particular compound, the route of administration, the severity of the condition being treated, and the condition and past history of the patient being treated.

In the present application, a "therapeutically and/or prophylactically effective amount" of a medicament herein for preventing and/or treating tuberculosis refers to a sufficient amount of the medicament to prevent and/or treat tuberculosis at a reasonable effect/risk ratio for any medical treatment and/or prophylaxis to treat a disorder.

The total daily amount of the medicament for preventing and/or treating tuberculosis of the present application must be decided by a physician in charge of the medical judgment within the scope of the reliable medical judgment. For any particular patient, the particular therapeutically effective dose level will depend on a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific drug used for preventing and/or treating tuberculosis; the specific composition employed; age, weight, general health, sex and diet of the patient; the administration time, administration route and excretion rate of the specific drug for preventing and/or treating tuberculosis are adopted; duration of treatment; a drug for preventing and/or treating tuberculosis used in combination with or simultaneously with the specific drug for preventing and/or treating tuberculosis; and similar factors well known in the medical arts. It is common practice in the art to start doses of the drug for the prevention and/or treatment of tuberculosis from below the level required to obtain the desired therapeutic effect and to gradually increase the dose until the desired effect is obtained.

In some embodiments, the content of indinavir and/or derivative thereof in the kit is formulated according to the following daily dosage criteria:

indinavir and/or derivatives thereof: 0.1mg/kg body weight/day to 100mg/kg body weight/day.

indinavir and/or derivatives thereof: 0.1mg/kg body weight/day to 10mg/kg body weight/day.

The dosage of indinavir and/or its derivatives described in this example is calculated as the amount of compound present in the unit dosage form.

In some embodiments, the indinavir and/or derivative thereof is present in the unit dosage form in an amount of from 1mg to 5000mg.

In some embodiments, the indinavir and/or derivative thereof is present in the unit dosage form in an amount of from 10mg to 500mg.

In some embodiments, the indinavir and/or derivative thereof is present in the unit dosage form in an amount of from 20mg to 300mg.

The method for differentiating THP-1 macrophages in the examples is as follows:

THP-1 (Tohoku Hospital Pediatrics-1) cells (human monocytic leukemia cells) were purchased from the national academy of sciences cell bank and cultured in 1640 medium containing 10% fetal bovine serum, 0.05mM beta-mercaptoethanol, 10mM HEPES and 1mM sodium pyruvate at 37℃with 5% CO ₂ Is cultured in a cell culture chamber of (2) in a 96-well plate at a ratio of 2.5X10 per well ⁴ The number of cells was plated, stimulated overnight with PMA (phorbol ester) at a final concentration of 50ng/ml, allowed to differentiate into macrophages, and 24h later replaced with the 1640 medium described above.

Example 1

This example shows the effect of various concentrations of indinavir on THP-1 macrophage survival.

Determination of cell viability: after the induced differentiated macrophages are continuously cultured for 24 hours, indinavir is added, the concentration of the indinavir is 5 mu M, 10 mu M and 20 mu M respectively, three repetitions are respectively carried out on each concentration, solvent DMSO is used as a control hole, a cell-free culture medium is used as a blank hole, the temperature is 37 ℃, and the concentration of CO is 5 percent ₂ And (3) continuing culturing for 48 hours, detecting the activity of the cells by using a CCK-8 kit, adding 10 mu l of CCK-8 into each hole, culturing for 2 hours in an incubator, measuring the absorbance at OD 450nm by using an enzyme-labeled instrument, and calculating the survival rate of the cells.

Cell viability was calculated as follows:

cell viability = [ (As-Ab)/(Ac-Ab) ]x100%;

as: experimental wells (cell-containing medium, CCK-8, indinavir at different concentrations);

ac: control wells (cell-containing medium, CCK-8, DMSO);

ab: blank wells (cell-free medium, CCK-8).

Fig. 1 is a dose response curve of indinavir, and the results show that there is no significant difference in the survival rate of THP-1 macrophages after treatment with different indinavir in the experimental group compared with the control group, and it is seen that indinavir has lower toxicity to THP-1.

Example 2

This example is the use of indinavir in inhibiting the growth of mycobacterium tuberculosis.

The application of the embodiment comprises the following steps:

s1, infection of mycobacterium tuberculosis: THP-1 cells were 2.5X10 per well in 24 well plates ⁵ The number of individual cells was induced to macrophages as described above, 25. Mu.M indinavir was added to the experimental group, the same volume of solvent DMSO was added to the control group, after pretreatment for 1H, cells were infected with Mycobacterium tuberculosis H37Rv at MOI (multiplicity of infection ) =10, washed 3 times with PBS (phosphate buffer saline, phosphate buffered saline) after 4H, and incubated at 37℃with 5% CO in 1640 complete medium ₂ The culture was continued in the incubator of (2) during which the addition of 25. Mu.M indinavir in the experimental group and the control group of equal volumes of solvent DMSO was maintained.

S2, plating count CFU (Colony-Forming Units): after 4h and 72h of infection, the cells were lysed with 0.025% SDS lysate (SDS Lysis Buffer) at 10 respectively ² 、10 ³ Dilution plating, bacterial incubator culture at 37 ℃ for about three weeks to count CFU.

In this example, the inhibition of 50 μm indinavir on the level of THP-1 macrophages against mycobacterium tuberculosis H37Rv is shown in fig. 2, and ns in fig. 2 represents no significant difference, and represents significantly less than 0.0001, and the result shows that indinavir can inhibit the growth of mycobacterium tuberculosis in macrophages, and compared with DMSO control group, the survival ability of mycobacterium tuberculosis is obviously reduced after indinavir is added, and the survival rate is reduced by 45-59%.

Example 3

This example is a Minimum Inhibitory Concentration (MIC) test of indinavir.

The detection steps in this embodiment are as follows:

taking a 96-well plate, adding 100 mu L of ddH into each well at the outermost periphery of the 96-well plate ₂ 98. Mu.L of 7H9-OADC complete medium was added to each well of O, B2-G2, and 50. Mu.L of 7H9-OADC complete medium was added to the remaining wells. 2. Mu.L of indinavir was added per well of B2-D2 and 2. Mu.L of INH (isoniazid) was added per well of E2-G2 as positive control. Double dilution from B2-G2 to B10-G10, B11-G11 was negative control wells without drug treatment, and finally 2X 10-containing wells (except ddH2O wells) were added to each well ⁵ CFU/50 mu L of Mycobacterium tuberculosis H37Rv bacterial liquid is placed in a bacteria incubator at 37 ℃ for 10-14 days after being sealed by a sealing film. After removal, bacterial inhibition was observed with different concentrations of drug and MIC values were read.

Table 1 shows that indinavir and INH have MIC for Mycobacterium tuberculosis H37Rv, and the result shows that the minimum inhibitory concentration of indinavir for Mycobacterium tuberculosis in vitro is more than 500 mu M and far more than the cell level use concentration, and the antituberculosis effect of indinavir at macrophage level is not directly acted on bacteria as antibiotics, but is used for inhibiting the intracellular survival of tuberculosis by regulating the host immune response through a target host. The minimum inhibitory concentration of INH on Mycobacterium tuberculosis is 0.02 mug/mL, which is consistent with the concentration range of the related technology, and the reliability of the experimental data is demonstrated as a positive control.

Table 1 shows the MIC of indinavir and INH for Mycobacterium tuberculosis H37Rv

The present application is not limited to the above embodiment. The above embodiments are merely examples, and embodiments having substantially the same configuration and the same effects as those of the technical idea within the scope of the present application are included in the technical scope of the present application. Further, various modifications that can be made to the embodiments and other modes of combining some of the constituent elements in the embodiments, which are conceivable to those skilled in the art, are also included in the scope of the present application within the scope not departing from the gist of the present application.

Claims

1. Use of indinavir and/or its derivatives in the preparation of antituberculosis drugs.

2. The use according to claim 1, wherein the derivative of indinavir comprises a pharmacologically acceptable salt thereof;

and/or the pharmacologically acceptable salts include salts with inorganic acids, organic acids, alkali metals, alkaline earth metals, and amino acids.

3. The use according to claim 1, wherein the antitubercular drug is for the prevention or treatment of infection by mycobacterium tuberculosis.

4. The use of claim 1, wherein the antitubercular drug is prepared from a material further comprising a pharmaceutically acceptable carrier;

and/or the pharmaceutically acceptable carrier comprises at least one of diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, sweeteners, and flavoring agents.

5. A medicament for preventing and/or treating tuberculosis, characterized in that the active ingredient of the medicament comprises indinavir and/or derivatives thereof.

6. A medicament for the prevention and/or treatment of tuberculosis according to claim 5, wherein the mass content of indinavir and/or its derivatives in the medicament is between 0.1% and 99%.

7. A medicament for the prophylaxis and/or treatment of tuberculosis according to claim 5, wherein the tuberculosis comprises tuberculosis or extrapulmonary tuberculosis.

8. A medicament for the prophylaxis and/or treatment of tuberculosis according to claim 5, wherein the medicament is administered in at least one selected from the group consisting of powder, tablet, granule, capsule, solution, emulsion and suspension.

9. A kit for the treatment of tuberculosis, characterized in that it comprises a medicament as described in any one of claims 5 to 7.

10. The tuberculosis treatment kit of claim 9, wherein the content of indinavir and/or derivatives thereof in the kit is formulated according to the following daily dosage criteria: